5HQ8

Co-crystal Structure of human SMYD3 with a MEKK2 peptide at 2.13A

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-D
Synchrotron site	APS
Beamline	21-ID-D
Temperature [K]	100
Detector technology	CCD
Collection date	2014-11-12
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	1.07808
Spacegroup name	P 1 21 1
Unit cell lengths	53.109, 118.155, 84.782
Unit cell angles	90.00, 90.61, 90.00

Refinement procedure

Resolution	25.430 - 1.720
R-factor	0.1755
Rwork	0.174
R-free	0.20110
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.018
RMSD bond angle	1.419
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHENIX
Refinement software	PHENIX (dev_1801)

Data quality characteristics

	Overall
Low resolution limit [Å]	50.000
High resolution limit [Å]	1.592
Number of reflections	138107
<I/σ(I)>	16.11
Completeness [%]	99.8
Redundancy	3.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7	295	Human SMYD3 (1-428) with SAM in the SAM-binding site was concentrated to 11.7 mg/mL in 25 mM Tris-HCl pH 8 and 150 mM NaCl. Peptide (21st Century Biochemical) was solubilized in 100mM water. Two ul peptide was added to 200 ul of protein (five fold molar excess of peptide) and allowed to complex with the protein overnight at 4C. Crystals were obtained from a Hampton Index HT broad screen in sitting drops at 22C with condition H7 (0.15 DL-malic acid pH 7.0, 20% w/v PEG 3,350). Crystals were flash frozen in 20 percent ethylene glycol in well solution prior to data collection